title: Transorganellar complementation redefines the biochemical continuity of endoplasmic reticulum and chloroplasts.
abstract: Tocopherols are nonpolar compounds synthesized and localized in plastids but whose genetic elimination specifically impacts fatty acid desaturation in the endoplasmic reticulum (ER), suggesting a direct interaction with ER-resident enzymes. To functionally probe for such interactions, we developed transorganellar complementation, where mutated pathway activities in one organelle are experimentally tested for substrate accessibility and complementation by active enzymes retargeted to a companion organelle. Mutations disrupting three plastid-resident activities in tocopherol and carotenoid synthesis were complemented from the ER in this fashion, demonstrating transorganellar access to at least seven nonpolar, plastid envelope-localized substrates from the lumen of the ER, likely through plastid:ER membrane interaction domains. The ability of enzymes in either organelle to access shared, nonpolar plastid metabolite pools redefines our understanding of the biochemical continuity of the ER and chloroplast with profound implications for the integration and regulation of organelle-spanning pathways that synthesize nonpolar metabolites in plants.
authors.affiliation: Departments of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI 48824, USA.
authors.forename: Payam
authors.initials: P
authors.lastname: Mehrshahi
authors.forename: Giovanni
authors.initials: G
authors.lastname: Stefano
authors.forename: Joshua Michael
authors.initials: JM
authors.lastname: Andaloro
authors.forename: Federica
authors.initials: F
authors.lastname: Brandizzi
authors.forename: John E
authors.initials: JE
authors.lastname: Froehlich
authors.forename: Dean
authors.initials: D
authors.lastname: DellaPenna
mesh_terms: D017360:Arabidopsis; D053898:Biosynthetic Pathways / Q000235:genetics*; D002338:Carotenoids / Q000096:biosynthesis / Q000378:metabolism; D002736:Chloroplasts / Q000378:metabolism*; D003001:Cloning, Molecular; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D004798:Enzymes / Q000378:metabolism*; D005816:Genetic Complementation Test / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D024505:Tocopherols / Q000378:metabolism*
chemical_list: D004798:Enzymes; D002338:Carotenoids; D024505:Tocopherols
keywords: MAM; PLAM; hemifusion; metabolism; vitamin E